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M200868200v1
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Papers In Press, published online ahead of print May 24, 2002
J. Biol. Chem, 10.1074/jbc.M200868200
Submitted on January 28, 2002
Revised on April 14, 2002
Accepted on May 24, 2002

Accelerated phagocytosis of amyloid-beta by mouse and human microglia overexpressing the M-CSF receptor

Olivera M. Mitrasinovic and Greer M. Murphy Jr

Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, CA 94305-5485

Corresponding Author: gmurphy{at}leland.stanford.edu

Microglia surrounding Abeta plaques in Alzheimer's disease (AD) and in the APPV717F transgenic mouse model of AD have enhanced immunoreactivity for the macrophage colony stimulating factor receptor (M-CSFR), encoded by the protooncogene c-fms. Increased expression of M-CSFR on cultured microglia results in proliferation and release of the pro-inflammatory cytokines and expression of inducible nitric oxide synthase. We transfected mouse BV-2 and human SV-A3 microglia to overexpress M-CSFR and examined microglial phagocytosis of fluorescein-conjugated Abeta . Flow cytometry and laser confocal microscopy showed accelerated phagocytosis of Abeta in mouse and human microglia due to M-CSFR overexpresssion that was time and concentration dependent. In contrast, microglial uptake of one micrometer diameter polystyrene microspheres was not enhanced by M-CSFR overexpression. Microglial uptake of Abeta was blocked by cytochalasin D, which inhibits phagocytosis. M-CSFR overexpression increased the mRNA for macrophage scavenger receptor A, and fucoidan blocking of macrophage scavenger receptors inhibited uptake of Abeta . M-CSFR antibody blocking experiments demonstrated that increased Abeta uptake depended on the interaction of the M-CSFR with its ligand. These results suggest that overexpression of M-CSFR in APPV717F mice may prime microglia for phagocytosis of Abeta after immunization.


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